T4 DNA ligase

Product Information
T4 DNA ligase can catalyze the reaction where the 5 '-P and 3' -OH ends of adjacent DNA strands are bound by phosphodiester bonds. It requires ATP as a coenzyme and can not only catalyze the connection of DNA between sticky ends or smooth ends, but also catalyze DNA.
Connections with DNA and a few Rnas. It can be applied to join double-stranded DNA molecules at sticky ends or flat ends, and can also be used to repair gaps on double-stranded DNA, RNA or DNA-RNA hybrids.
Reaction buffer: 500 mM Tris-HCl, pH 7.5, 50 mM MgCl₂, 25 mM DTT, 5 mM ATP
Instructions Prepare the reaction system according to the following components:
2 µL reaction buffer; 50 ng (0.2-1 pmol) carrier; Insert fragments of 1-50 ng (0.2-10 pmol); 1 µL T4 DNA Ligase; Add ddH2O to complete up to 20 µL;
When connecting two fragments, the molar ratio of the carrier to the inserted fragment is usually 1:3 ideal experimental results can be obtained within the range of 1:1 to 1:10). The molar concentration is calculated based on the size and concentration of the fragment during extraction.
2) If it is a sticky end connection reaction, connect at 22°C (16-37°C) for 30 minutes, and the connection product can be directly used for transformation (or frozen storage at -20°C).

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.